Rescue and repair of stalled ribosome damaged by ribosome-specific ribotoxins

被核糖体特异性核毒素损坏的停滞核糖体的拯救和修复

• 批准号: 10799097
• 负责人: Raven H Huang
• 金额: $ 5.95万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10799097
• 关键词: Antidotes; Attenuated; Bacteria; Biochemical; Biochemistry; Bioinformatics; Biological Process; Cell Death; Cells; Coupled; Data; Event; In Vitro; Microbiology; Prevention; Proteins; RNA; Ribosomal RNA; Ribosomes; System; Toxic effect; Translations; biological systems; cell killing; colicin; human disease; in vivo; insight; pathogenic bacteria; repaired; small molecule inhibitor; structural biology

Project Summary/Abstract Colicin E3 was the first ribotoxin to be characterized 50 years ago. It makes a single but precise cut of 16S rRNA in the decoding center of bacterial ribosome, resulting in stalled ribosome and eventual cell death. Over the last half of a century, it is unclear whether there exists a biological system that is able to reverse the ribosomal damage by colicin E3 to allow cell to survive. Employing approaches of bioinformatics, biochemistry, structural biology, and microbiology, we have uncovered a bacterial two- component system, RtcB and PrfH, as the antidote of colicin E3. Specifically, bacterial PrfH recognizes the damaged and stalled ribosome and performs ribosomal rescue. This is followed by RtcB repairing the damaged 30S ribosomal subunit. The sequential events described above are supported by abundant preliminary data from both our in vitro and in vivo studies. In this application, we plan to significantly expand our preliminary studies to systematically characterize the rescue and repair of bacterial ribosome with specific damage in the decoding center with the following three main aims: (1) We will provide insight into bacterial PrfH recognizing and rescuing the damaged and stalled 70S ribosome in vitro; (2) We will biochemically and structurally characterize bacterial RtcB in vitro, with the emphasis of PrfH-coupled RtcB repairing the damaged 30S ribosomal subunit; and (3) We will elucidate in vivo biological functions of RtcB- PrfH using an in vivo attenuated RNA damage system we have developed.

项目总结/文摘